Mass spectral determination of skeletal/cardiac actin isoform ratios in cardiac muscle

Harold Robert (Bob) III Bergen, Katalin Ajtai, Thomas P Burghardt, Angelito I. Nepomuceno, David C. Muddiman

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Skeletal and cardiac muscle contains actin isoforms that vary by two juxtaposed amino acids and two amino acid substitutions (Met299Leu and Ser358Thr). This close sequence homology does not allow cardiac and skeletal actin isoforms to be resolved in traditional SDS-PAGE analysis as the molecular weights (Δmass = 32Da) are not significantly different and the pIs are identical (5.2). Although cardiac actin is the predominant form in cardiac muscle, there appears to be a specific skeletal/cardiac actin ratio in a normal heart that may vary in a compromised or diseased heart. In an effort to ascertain the validity of this hypothesis we developed a mass spectrometric technique to measure the ratio of skeletal to cardiac actin. The technique involves purification of muscle actin and subsequent liquid chromatography coupled with electrospray ionization Fourier transform ion cylcotron resonance (LC/FTICR-MS) mass spectrometry. A 7 Tesla FTICR mass spectrometer was utilized to compare skeletal/cardiac actin isoform ratios. Additionally, a new dual electrospray ionization source was employed to determine accurate masses of the α-skeletal and α-cardiac actins.

Original languageEnglish (US)
Pages (from-to)1467-1471
Number of pages5
JournalRapid Communications in Mass Spectrometry
Volume17
Issue number13
DOIs
StatePublished - 2003

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Muscle
Actins
Protein Isoforms
Electrospray ionization
Amino Acids
Liquid chromatography
Mass spectrometers
Ion sources
Purification
Mass spectrometry
Fourier transforms
Substitution reactions
Molecular weight
Ions

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy

Cite this

Mass spectral determination of skeletal/cardiac actin isoform ratios in cardiac muscle. / Bergen, Harold Robert (Bob) III; Ajtai, Katalin; Burghardt, Thomas P; Nepomuceno, Angelito I.; Muddiman, David C.

In: Rapid Communications in Mass Spectrometry, Vol. 17, No. 13, 2003, p. 1467-1471.

Research output: Contribution to journalArticle

Bergen, Harold Robert (Bob) III ; Ajtai, Katalin ; Burghardt, Thomas P ; Nepomuceno, Angelito I. ; Muddiman, David C. / Mass spectral determination of skeletal/cardiac actin isoform ratios in cardiac muscle. In: Rapid Communications in Mass Spectrometry. 2003 ; Vol. 17, No. 13. pp. 1467-1471.
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